Simultaneous detection of four food borne bacterial pathogens by metal hydroxide immobilized multiplex PCR method

Abstract

Salmonella enterica, Vibrio parahaemolyticus, Escherichia coli and Listeria monocytogenes are human pathogenic bacteria which could be contaminated with many types of food, especially meat products. Testing of food for those pathogens has become routine practice all over the world. However, since the available traditional detection methods are time consuming and labor intensive, there is a need for rapid, sensitive, specific and cost effective detection technique. In this study, a rapid and sensitive single tube Multiplex PCR method for simultaneous detection of those four bacterial pathogens by amplifica tion of target genes gyrB of V. parahaemolyticus, invA of S. enterica, eaeA of E.coli and hly of L. monocy togene was established. Concentration of bacterial cells from food samples for DNA extraction is an other problem in testing of food pathogens by PCR techniques. Metal hydroxide immobilization was applied prior to DNA extraction and found to be effective step to overcome this problem.